DEVELOPMENT AND VALIDATION OF A MONOCLONAL-ANTIBODY ENZYME-IMMUNOASSAY FOR MEASURING PROGESTERONE IN SALIVA

A nonextraction, competitive, solid-phase enzyme immunoassay with a mo noclonal antibody was developed and validated for measuring progestero ne in saliva. The antibody was raised against 11alpha-hydroxyprogester one hemisuccinate-bovine serum albumin conjugate and was indirectly im mobilized to the walls of microtiter wells. The labeled analyte (proge sterone-horseradish peroxidase conjugate) was homologous with the immu nogen. The lower detection limit (concentration equivalent to B0 - 3 S D) was 38 pmol/L of saliva sample. We validated the assay with studies to establish the independence of the concentration determined from th e volume of saliva assayed, quantitative recovery of progesterone adde d to saliva, interference from possible cross-reactants, and agreement with a similar assay that incorporated an extraction step. In additio n, we determined luteal-phase concentrations of salivary progesterone in normal women and compared the results with those of an older assay involving a polyclonal antibody.